Ionomycin ameliorates hypophosphatasia via rescuing alkaline phosphatase deficiency-mediated L-type Ca^(2+) channel internalization in mesenchymal stem cells

The loss-of-function mutations in the ALPL result in hypophosphatasia(HPP), an inborn metabolic disorder that causes skeletal mineralization defects. In adults, the main clinical features are early loss of primary or secondary teeth, osteoporosis, bone pain,chondrocalcinosis, and fractures. However, guidelines for the treatment of adults with HPP are not available. Here, we show that ALPL deficiency caused a reduction in intracellular Ca2+ influx, resulting in an osteoporotic phenotype due to downregulated osteogenic differentiation and upregulated adipogenic differentiation in both human and mouse bone marrow mesenchymal stem cells(BMSCs). Increasing the intracellular level of calcium in BMSCs by ionomycin treatment rescued the osteoporotic phenotype in alpl+/- mice and BMSC-specific(Prrx1-alpl-/-) conditional alpl knockout mice. Mechanistically, ALPL was found to be required for the maintenance of intracellular Ca2+ influx, which it achieves by regulating L-type Ca2+ channel trafficking via binding to the α2δsubunits to regulate the internalization of the L-type Ca2+ channel. Decreased Ca2+ flux inactivates the Akt/GSK3β/β-catenin signaling pathway, which regulates lineage differentiation of BMSCs. This study identifies a previously unknown role of the ectoenzyme ALPL in the maintenance of calcium channel trafficking to regulate stem cell lineage differentiation and bone homeostasis. Accelerating Ca2+ flux through L-type Ca2+ channels by ionomycin treatment may be a promising therapeutic approach for adult patients with HPP.

EXTERNALIZATION AND INTERNALIZATION OF CARDIAC ENDOTHELIN RECEPTORS DURING DIFFERENT PHASES OF SEPSIS IN RAT

To study the redistribution of endothelin- 1 (ET- 1) receptors in two subcellular organelles , the sarcolemmal membrane and the light vesicle, of rat heart during the progression of sepsis. Methods. Sepsis was induced by cecal ligation and puncture (CLP). ET1 receptor was assayed by using [125I]- ET1 binding. Marker enzyme activities, protein yield, and dry- to- wet weight ratio of cardiac membranes were measured. Results. Septic rat heart exhibited two distinct phases: an initial hyperdynamic phase( 9h after CLP; early stage of sepsis) followed by a hypodynamic (18h after CLP, late stage of sepsis) phase. [125I]- ET1 binding study showed that during early stage of sepsis, the Bmax of ET1 receptors was increased by 30％ in sarcolemma but decreased by 19％ in light vesicles, while during late stage of sepsis, the Bmax was decreased by 24％ in sarcolemma but increased by 38％ in light vesicles.The total binding of sarcolemma and light vesicles was increased by 25％ during early stage of sepsis but decreased by 17％ during late stage of sepsis. Conclusions. These data indicated that ET1 receptors in the rat heart were externalized from light vesicles to sarcolemmal membranes during early hyperdynamic phase while internalized from surface membranes to intracellular compartment during late hypodynamic phase of sepsis.

Intracellular distribution and internalization pathways of guanidinylated bioresponsive poly(amido amine)s in gene delivery

Guanidinylated bioresponsive poly(amido amine)s polymers, CAR-CBA and CHL-CBA, weresynthesized by Michael-type addition reaction between guanidine hydrochloride(CAR) orchlorhexidine(CHL) and N,N-cystaminebisacrylamide(CBA). Previous studies have shownthat both polymers had high transfection efficiencies as gene delivery carriers. In this study,we investigated the nucleolus localization abilities and cellular internalization pathways ofthese two polymers in gene delivery. Each polymer condensed plasmid DNA(p DNA) andformed nanoparticle complexes, and then their transfection studies were performed inMCF-7 cells. Both complexes were found enriched in nucleolus after cellular transfection,and their transfection efficiencies were significantly improved when transfection was per-formed on MCF-7 cells arrested at M phase. The transfection efficiency of CAR-CBA-pDNAwas inhibited by chlorpromazine, and cell endosomes were disrupted after being exposedto CAR-CBA-pDNA. In regards to CHL-CBA-pDNA, its transfection efficiency was not affected by three types of endocytosis inhibitors used in the study, and CHL-CBA-pDNA showed no effect on endosomes. Cellular lactate dehydrogenase release and membrane morphology were changed after cells were transfected by the two complexes. The results indicated that both CAR-CBA and CHL-CBA polymers demonstrated good nucleolus localization abilities. It was beneficial for transfection when cells were arrested at M phase. CAR-CBA-pDNA cellular internalization was involved with clathrin-mediated endocytosis pathway, and escaping from endosomal entrapment, while the cellular uptake of CHL-CBA-pDNA occurs via clathrin-and caveolae-independent mechanism.

Follicle-stimulating Hormone(FSH) Induced Internalization of Porcine FSH Receptor in Cultured Porcine Granulosa Cells and Chinese Hamster Ovary Cells Transfected with Recombinant Porcine FSH Receptor cDNA

In order to study the fate of human follicle-stimulating hormone (FSH) when hormone binds to its receptor, a quick biochemical method that can differentiate between the surface-bound and internalized hormone was used to determine the internalization induced by FSH in cultured both porcine granulosa cells and Chinese hamster ovary (CHO) cells expressing recombinant porcine FSH receptor. The results showed that FSH was slowly internalized, and the internalized radioactivity (acid resistant) reached a peak 10-12 h after addition of 125 I-hFSH. It was suggested that FSHR do not get internalized rapidly under physiological circumstances precisely because the appropriate sequences are absent.

GPCR/endocytosis/ERK signaling/S2R is involved in the regulation of the internalization,mitochondria-targeting and-activating properties of human salivary histatin 1

Human salivary histatin 1(Hst1)exhibits a series of cell-activating properties,such as promoting cell spreading,migration,and metabolic activity.We recently have shown that fluorescently labeled Hst1(F-Hst1)targets and activates mitochondria,presenting an important molecular mechanism.However,its regulating signaling pathways remain to be elucidated.We investigated the influence of specific inhibitors of G protein-coupled receptors(GPCR),endocytosis pathways,extracellular signal-regulated kinases1/2(ERK1/2)signaling,p38 signaling,mitochondrial respiration and Na+/K+-ATPase activity on the uptake,mitochondria-targeting and-activating properties of F-Hst1.We performed a si RNA knockdown(KD)to assess the effect of Sigma-2 receptor(S2R)/Transmembrane Protein 97(TMEM97)—a recently identified target protein of Hst1.We also adopted live cell imaging to monitor the whole intracellular trafficking process of F-Hst1.Our results showed that the inhibition of cellular respiration hindered the internalization of F-Hst1.The inhibitors of GPCR,ERK1/2,phagocytosis,and clathrin-mediated endocytosis(CME)as well as siRNA KD of S2R/TMEM97 significantly reduced the uptake,which was accompanied by the nullification of the promoting effect of F-Hst1 on cell metabolic activity.Only the inhibitor of CME and KD of S2R/TMEM97 significantly compromised the mitochondria-targeting of Hst1.We further showed the intracellular trafficking and targeting process of F-Hst1,in which early endosome plays an important role.Overall,phagocytosis,CME,GPCR,ERK signaling,and S2R/TMEM97 are involved in the internalization of Hst1,while only CME and S2R/TMEM97 are critical for its subcellular targeting.The inhibition of either internalization or mitochondria-targeting of Hst1 could significantly compromise its mitochondria-activating property.

Body-focused Anxiety in Women: Associations with Internalization of the Thin-ideal, Dieting Frequency, Body Mass Index and Media Effects

Exposure to media that portrays thin women as ideal and attractive can lead to women internalizing the thin ideal, which results in incorporating societal standards of thinness into belief systems. Internalization of the thin-ideal is associated with numerous detrimental effects on women, including decreased levels of self-esteem and increased levels of body-focused anxiety, negative emotions and disordered eating. The present study utilized a sample of women (N = 208) aged between 18 and 67 years (M = 29.44, SD = 13.08) to examine the relationship between internalization of the thin- ideal, body-focused anxiety, body mass index (BMI), and dieting frequency. Correlational, regression and mediation analyses conducted on the data showed that internalization of the thin-ideal, BMI and dieting frequency significantly contributed to body-focused anxiety in women. In addition, body-focused anxiety fully mediated the relationship between internalization of the thin-ideal and dieting frequency among women. BMI did not moderate the relationship between internalization of the thin-ideal and body-focused, indicating that women who internalize the thin-ideal are less vulnerable to dieting unless experiencing body-focused anxiety. The results of the current study enhance our understanding of the relationship between internalization of the thin-ideal, body-focused anxiety, BMI, and dieting frequency among women. Clinical implications will be discussed.

＂Half-a-Day＂ Syndrome： A Mode of Internalization of Vocational and Technical Education by the Youths in the 20th Century Nigeria

This research focused on ＂half-a-day＂ as a mode of internalizing vocational and technical education by the youths in Nigeria. It traced the origin of ＂half-a-day＂ linking it with the ingenuity of the youths to make the formal education that they have acquired operational. It also discussed the traditional education with its apprenticeship system as being modified through the ＂half-a-day＂ syndrome. The study is historical and descriptive therefore, social survey method was employed using unscheduled interview and consulting relevant literature. It was discovered that, apart from the failure of the educational system due to lack of proper infrastructure, the youths, especially those from the lower class, have reasoned that the salaries from the white-collar jobs may not be enough to sustain them. It is, therefore, recommended, among others, that： Government can institutionalise the ＂half-a-day＂ as a form of summer school and certify both the centres of learning and learners.

The Observation of FSH’s Cellular Internalization

Follicle stimulating hormone (FSH) is a kind of glycoprotein gonadotropin, and plays an important role in the diagnosis and treatment of infertility. Follicle stimulating hormone receptor (FSHR) is a kind of G protein coupled receptor (GPCR), found in the ovary and testes, and its activation is required for the hormonal operation during the breeding period. In this study, an experimental model of FSHR mediated FSH into cell membrane, which exhibited a phenomenon of fluorescent localized on cell surfaces internalized into cell interior, was established to verify biological activity of FSH.

On the necessity and countermeasures of the ＂internalization＂ construction of the China-characterized public diplomacy under the new situations

In the era of the globalization of the economy, the network of the information, and the socialization of the citizens, there is the tendency that a country＇s domestic problems will be internationalized, while the international issues will be domestic. Combined with the background of China to realize the ＂China Dreams＂, in the field of the public diplomacy, we should pay sufficient attention to the ＂internalization＂ construction in the aspects of the value ideas, the mode of the systems, the political propositions, and the cultural influence and so on, which is the benign interaction involving the governments, the citizens and the social organizations. In this paper, the author casts a brick to attract jade. Through the discussion of the necessity of the ＂internalization＂ construction of the public diplomacy, the author carries out the discussion from the social and the cultural dimensions, in order to arouse the attention to the ＂internalization＂ construction of the public diplomacy, so that our country will be more influential in the politics, and more approachable in the images, and more appealing in the morality.

Research for Competitiveness Enhancement Problem of Regional Industry Based on Brand Internalization Theory

Along with the quick development of social economy, market competition has already entered into a new stage, namely brand competition era. This paper is to put forward scientifically rational countermeasures and suggestions through taking brand internalization theory as basis and starting from regional industry competitiveness. Thus it would increase regional industry competitiveness of brand.

The role repositioning of teacher and student in the process of knowledge internalization under flipped classroom

Flipped classroom model led to the transformation of university teachers＇ teaching role. From the perspective of the sociology of knowledge, the teacher professional development is a process of transformation of the role of teachers, and role transformation and the way of actors involved in the knowledge are associated. Participating in knowledge in different ways, role types are also different. Thus, changing the way ofparticipating in knowledge is the basic way to participate in the transformation of the role. Classification of roles is not the classification ofhumans.A social may might play many roles of many types in his life andmay have the function of a variety of roles at the same time. Meanwhile, it can be transferred between characters, especially in the case of depending on the same knowledge system.As long as the ways of involvement are different,the functions and types of roles are also different. University teachers are those who play a variety of roles of knowledge and society, some being the teaching teachers of organization and dissemination of knowledge, some being scientific and teaching teachers who focus on organizing and disseminating knowledge as well as knowledge creation, and knowledge creation and heavy teacher teaching and research, while others are knowledge creation based research teachers. In the field of the sociology of knowledge, university teachers should be scientific teaching roles depending on a deep knowledge and participating in knowledge in a variety of ways. From the perspective of the sociology of knowledge, according to Eph · Znaniecki＇ s role classification, university teachers participate in knowledge in three ways： teaching, learning and researching. Teaching, learning and researching are relative but different ways of participating in knowledge. State they are relative because itshows teaching can promote learning, learning can promote researching, researching can also promote learning and teaching. The homochromous interaction of teaching, leaming and researching, knowledge is in constant state of development and generation.

CREG inhibits VSMCs proliferation by modulating the internalization of IGFII

Background To study the molecular mechanisms of CREG(the cellular repressor of E1A-stimulated gene) on proliferation of VSMCs in vitro.Methods The pRc/CMV-CREG plasmid or the pSM2-siCREG plasmid was transferred into human vascular smooth muscle cells(hVSMCs) to produce the cell clone that over-expression or down-expression of CREG respectively.BrdU assay and FACS cell cycle analysis were used to detect the proliferation of cells.Western blotting and immunocytochemistry show the expression and localization of IGF2R in hVSMCs.RT-PCR and ELISA assay determined the expression and secretion of IGFII factor. Alex488-labeled rhIGFII was used to investigate the endocysis of cells.And the blockade of IGFII internalization by treatment both the neutralized antibody of anti-IGF2R and rsIGF2R detected the effect of IGFII on VSMCs growth. Furthermore,Western blotting and signal pathway inhibitor were used to analysis the activation of PI3K/AKT and ERK on VSMCs proliferation.Results Western blotting identified that the expression of CREG in hVSMCs-CREG cells increased compared to control cell,and the decreased obviously in hVSMCs-siCREG cells.Meanwhile,the overexpression of CREG in cells was detected to inhibit the proliferation of VSMCs and to enhance the distribution of IGF2R in cellular membrane.Furthermore,overexpression of CREG also accelerated the endocysis of IGFII in hVSMCs-CREG,and attenuate the secretion of IGFII into cell medium by ELISA analysis and Alex488 labeled IGFII analysis.Blockade experiments both neutralized antibody of IGF2R and rhIGF2R fragment determined that enhancement of IGFII secretion promoted the VSMCs proliferation,and PI3K/AKT and ERK signal pathway mediated the effect of IGFII on VSMCs.Conclusions Altogether, these data indicate that CREG inhibits the proliferation of hVSMCs through interfering into the internalization pathway of IGF2R-IGFII.

Study on the Promoting Mechanism of Teachers'Ethics Internalization in Universities

On the basis of clarifying the teachers'Ethics internalization theory,this paper analyzes the typical dilemma in teachers'Ethics internalization.To resolve these problems,this paper construct an overall framework,including security mechanism,normative mechanism,education mechanism,evaluation mechanism and supervision mechanism,which the external conditions are used to constantly strengthen the moral needs of the subject,then enhance the consciousness of moral internalization,improve the effectiveness of teachers'moral construction.

Erratum to“Tumor Microenvironment-Specific Chemical Internalization for Enhanced Gene Therapy of Metastatic Breast Cancer”

In the Research Article “Tumor Microenvironment-Specific Chemical Internalization for Enhanced Gene Therapy of Metastatic Breast Cancer,” the authors made an error in Fig. 6A. In Fig. 6A, the image of “pre” in the “Free Cy5-siRNA” group was misused. When incorporating the final images in Fig. 6A, another image of “24 h” in the “Free Cy5-siRNA” group was inadvertently chosen as the “Pre” image, so they appear to be remarkably similar. After the error was discovered, the authors repeated the experiments that concluded that this error did not affect the results, discussion, or conclusion of this paper. Figure 6A has now been corrected in the PDF and HTML (full text).

Mechanism of internal thermal runaway propagation in blade batteries

Blade batteries are extensively used in electric vehicles,but unavoidable thermal runaway is an inherent threat to their safe use.This study experimentally investigated the mechanism underlying thermal runaway propagation within a blade battery by using a nail to trigger thermal runaway and thermocouples to track its propagation inside a cell.The results showed that the internal thermal runaway could propagate for up to 272 s,which is comparable to that of a traditional battery module.The velocity of the thermal runaway propagation fluctuated between 1 and 8 mm s^(-1),depending on both the electrolyte content and high-temperature gas diffusion.In the early stages of thermal runaway,the electrolyte participated in the reaction,which intensified the thermal runaway and accelerated its propagation.As the battery temperature increased,the electrolyte evaporated,which attenuated the acceleration effect.Gas diffusion affected thermal runaway propagation through both heat transfer and mass transfer.The experimental results indicated that gas diffusion accelerated the velocity of thermal runaway propagation by 36.84%.We used a 1D mathematical model and confirmed that convective heat transfer induced by gas diffusion increased the velocity of thermal runaway propagation by 5.46%-17.06%.Finally,the temperature rate curve was analyzed,and a three-stage mechanism for internal thermal runaway propagation was proposed.In Stage I,convective heat transfer from electrolyte evaporation locally increased the temperature to 100℃.In Stage II,solid heat transfer locally increases the temperature to trigger thermal runaway.In StageⅢ,thermal runaway sharply increases the local temperature.The proposed mechanism sheds light on the internal thermal runaway propagation of blade batteries and offers valuable insights into safety considerations for future design.

Endoscopic polidocanol foam sclerobanding for the treatment of grade Ⅱ-Ⅲ internal hemorrhoids:A prospective,multi-center,randomized study

BACKGROUND Endoscopic rubber band ligation(ERBL)is a nonsurgical technique for the treatment of symptomatic internal hemorrhoids but is limited by recurrence and post-procedural pain.AIM To evaluate satisfaction,long-term recurrence,and post-procedural pain in managing internal hemorrhoids using a combination of polidocanol foam sclerotherapy and ERBL.METHODS This was a prospective,multicenter,randomized study.A total of 195 consecutive patients diagnosed with grade II-III internal hemorrhoids were enrolled from four tertiary hospitals and randomly divided into a cap-assisted endoscopic polidocanol foam sclerobanding(EFSB)or an ERBL group.All patients were followed-up for 12 months.Symptom-based severity and post-procedural pain were assessed using a hemorrhoid severity score(HSS)and a visual analog scale(VAS).Continuous variables were reported as medians and interquartile range.RESULTS One hundred and ninety-five patients were enrolled,with 98 in the EFSB group.HSS was lower in the EFSB group than in the ERBL group at 8 weeks[4.0(3.0-5.0)vs 5.0(4.0-6.0),P=0.003]and 12-month[2.0(1.0-3.0)vs 3.0(2.0-3.0),P<0.001]of follow-up.The prolapse recurrence rate was lower in the EFSB group at 12 months(11.2%vs 21.6%,P=0.038).Multiple linear regression analysis demonstrated that EFSB treatment[B=-0.915,95%confidence interval(CI):−1.301 to−0.530,P=0.001]and rubber band number(B=0.843,95%CI:0.595-1.092,P<0.001)were negatively and independently associated with the VAS score 24 hours post-procedure.The median VAS was lower in the EFSB group than in the ERBL[2.0(1.0-3.0)vs 3.0(2.0-4.0),P<0.001].CONCLUSION Cap-assisted EFSB provided long-term satisfaction and effective relief from the recurrence of prolapse and pain 24 hours post-procedure.

Selective internal radiation therapy segmentectomy:A new minimally invasive curative option for primary liver malignancies?

Transarterial radioembolization or selective internal radiation therapy(SIRT)has emerged as a minimally invasive approach for the treatment of tumors.This percutaneous technique involves the local,intra-arterial delivery of radioactive microspheres directly into the tumor.Historically employed as a palliative measure for liver malignancies,SIRT has gained traction over the past decade as a potential curative option,mirroring the increasing role of radiation segmentectomy.The latest update of the BCLC hepatocellular carcinoma guidelines recognizes SIRT as an effective treatment modality comparable to other local ablative methods,particularly well-suited for patients where surgical resection or ablation is not feasible.Radiation segmentectomy is a more selective approach,aiming to deliver high-dose radiation to one to three specific hepatic segments,while minimizing damage to surrounding healthy tissue.Future research efforts in radiation segmentectomy should prioritize optimizing radiation dosimetry and refining the technique for super-selective administration of radiospheres within the designated hepatic segments.

Seeing beneath the surface:Harnessing point-of-care ultrasound for internal jugular vein evaluation

Point-of-care ultrasound(POCUS)of the internal jugular vein(IJV)offers a noninvasive means of estimating right atrial pressure(RAP),especially in cases where the inferior vena cava is inaccessible or unreliable due to conditions such as liver disease or abdominal surgery.While many clinicians are familiar with visually assessing jugular venous pressure through the internal jugular vein,this method lacks sensitivity.The utilization of POCUS significantly enhances the visualization of the vein,leading to a more accurate identification.It has been demonstrated that combining IJV POCUS with physical examination enhances the specificity of RAP estimation.This review aims to provide a comprehensive summary of the various sonographic techniques available for estimating RAP from the internal jugular vein,drawing upon existing data.

Utilization of Basalt Saw Mud as a Spherical Porous Functional Aggregate for the Preparation of Ordinary Structure Concrete

To promote the production and application of artificial aggregates,save natural sand resources and protect the ecological environment,we evaluated the feasibility of using spherical porous functional aggregates(SPFAs) formed by basalt saw mud under autoclave curing in ordinary structural concrete.In our work,two types of prewetted functional aggregates were taken as replacements for natural aggregates with different volume substitution rates(0%,5%,10%,15%,20%,25%,and 30%) in the preparation of ordinary structural concrete with water-to-binder ratios(W/B) of 0.48 and 0.33.The effects of the functional aggregate properties and content,W/B,and curing age on the fluidity,density,mechanical properties and autogenous shrinkage of ordinary concrete were analyzed.The experimental results showed that the density of concrete declined at a rate of not more than 5%,and the 28 d compressive strength could reach 31.0-68.2 MPa.Low W/B,long curing age and high-quality functional aggregates were conducive to enhancing the mechanical properties of SPFAs concrete.Through the rolling effects,SPFAs can optimize the particle gradation of aggregate systems and improve the fluidity of concrete,and the water stored inside SPFAs provides an internal curing effect,which prolongs the cement hydration process and considerably reduces the autogenous shrinkage of concrete.SPFAs exhibits high strength and high density,as well as being more cost-effective and ecological,and is expected to be widely employed in ordinary structural concrete.